|Title:||Development of cell surface shaving proteomics for application in staphylococcal species|
|Publisher:||University of Sydney.|
Faculty of Science.
School of Molecular Bioscience.
|Abstract:||Staphylococci are important Gram-positive organisms that have close interactions with humans in the form of commensalism and pathogenicity. In particular, Staphylococcus aureus and Staphylococcus epidermidis cause high morbidity and mortality and are responsible for a high cost to healthcare systems worldwide. These species are known to have high degrees of multidrug resistance, biofilm formation capabilities and several virulence factors that allow for infection of human tissues. Surface-exposed proteins are the major biomolecules that are involved in environmental recognition, adhesion, signal transduction and initiation of colonization, but also represent one of the most difficult classes of proteins to analyse by proteomic methodologies. Here we review the literature for current proteomic techniques employed to characterise surface-exposed proteins and introduce the principle of cell surface shaving. Cell shaving was initially applied to S. aureus with a novel control strategy as a proof-of-principle to demonstrate the applicability of profiling surface proteomes (surfaceomes). Cell shaving was then applied to a S. aureus model of hyperantibiotic resistance to investigate the responses of this pathogen to extended antibiotic exposure. This also included the development of a new analysis method for cell shaving utilising hypergeometric statistics and Bayesian inference to calculate the probability of a protein being surface-exposed. This resulted in the identification of one of the largest surface-exposed proteins profiles in the literature for this organism and proteins associated with antibiotic stress and osmotolerance. Finally, a comparative profiling of two S. epidermidis strains was performed where a biofilm-forming and pathogenic strain was contrasted against a non-biofilm-forming, non-pathogenic strain and surface-exposed proteins associated with biofilm formation were catalogued. This represents the first surfaceome profile in S. epidermidis.|
|Access Level:||Access is restricted to staff and students of the University of Sydney . UniKey credentials are required. Non university access may be obtained by visiting the University of Sydney Library.|
|Rights and Permissions:||The author retains copyright of this thesis. It may only be used for the purposes of research and study. It must not be used for any other purposes and may not be transmitted or shared with others without prior permission.|
|Type of Work:||PhD Doctorate|
|Type of Publication:||Doctor of Philosophy Ph.D.|
|Appears in Collections:||Sydney Digital Theses (University of Sydney Access only)|
|solis_ns_thesis.pdf||PhD Thesis||3.36 MB||Adobe PDF|
|chapter2_supp_data_1.pptx||Supplementary data||599.99 kB||Microsoft Powerpoint|
|chapter2_supp_data_2.pdf||Supplementary data||189.26 kB||Adobe PDF|
|chapter2_supp_data_3.pdf||Supplementary data||318.06 kB||Adobe PDF|
|chapter3_supp_data_1.xlsx||Supplementary data||390.62 kB||Microsoft Excel|
|chapter3_supp_data_2.xlsx||Supplementary data||191.39 kB||Microsoft Excel|
|chapter3_supp_data_3.pptx||Supplementary data||1.9 MB||Microsoft Powerpoint|
|chapter4_supp_data_1.xlsx||Supplementary data||162.07 kB||Microsoft Excel|
Items in Sydney eScholarship Repository are protected by copyright, with all rights reserved, unless otherwise indicated.